Publication detail

Pyrosequencing on a glass surface

ALMEIDA, A. MANZ, A. NEUŽIL, P.

Original Title

Pyrosequencing on a glass surface

Type

journal article in Web of Science

Language

English

Original Abstract

We demonstrate the use of open-surface microfluidics to sequence DNA by pyrosequencing at the plain hydrophobically coated surface of a microscope glass cover slip. This method offers significant advantages in terms of instrument size, simplicity, disposability, and functional integration, particularly when combined with the broad and flexible capabilities of open-surface microfluidics. The DNA was incubated on superparamagnetic particles and placed on a hydrophobically coated glass substrate. The particles with bound DNA were moved using magnetic force through microliter-sized droplets covered with mineral oil to prevent water evaporation from the droplets. These droplets served as reaction “stations” performing pyrosequencing as well as washing stations. The resequencing protocol with 34-mer single-stranded DNA (ssDNA) was used to determine the reaction performance. The de novo sequencing was performed with 51-mer and 81-mer ssDNA. The method can be integrated with previously shown sample preparation and PCR into a single sample-to-answer system on a plain glass surface.

Keywords

DNA sequence determination; pyrosequencing; droplets; magnetic particles; open-surface microfluidics; virtual reaction chambers

Authors

ALMEIDA, A.; MANZ, A.; NEUŽIL, P.

Released

16. 2. 2016

Publisher

Royal Society of Chemistry

Location

Londyn

ISBN

1473-0197

Periodical

LAB ON A CHIP

Year of study

16

Number

6

State

United Kingdom of Great Britain and Northern Ireland

Pages from

1063

Pages to

1071

Pages count

9

URL

https://pubs.rsc.org/en/content/articlelanding/2016/LC/C6LC00114A

Full text in the Digital Library

http://hdl.handle.net/11012/203127

BibTex

@article{BUT122201,
  author="Ana V. {Almeida} and Andreas {Manz} and Pavel {Neužil}",
  title="Pyrosequencing on a glass surface",
  journal="LAB ON A CHIP",
  year="2016",
  volume="16",
  number="6",
  pages="1063--1071",
  doi="10.1039/c6lc00114a",
  issn="1473-0197",
  url="https://pubs.rsc.org/en/content/articlelanding/2016/LC/C6LC00114A"
}